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Urinary follicle stimulating hormone with high purity and preparation method thereof

A follicle-stimulating hormone, high-purity technology, applied in the field of high-purity follicle-stimulating hormone and its preparation, can solve problems such as side effects, FSH specific activity and purity are not very ideal

Active Publication Date: 2013-03-27
SHANGHAI TECHWELL BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The current preparation method of urine-derived FSH is mainly based on low-purity urinary follicle-stimulating hormone or menopausal gonadotropin (HMG) as the starting material, through hydrophobic chromatography, or monoclonal antibody immunochromatography and reversed-phase HPLC. Purification, but the specific activity and purity of the obtained FSH are not very satisfactory, and there are occasional reports of side reactions

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  • Urinary follicle stimulating hormone with high purity and preparation method thereof

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preparation example Construction

[0052] The preparation method of the high-purity urogenous FSH provided by the invention comprises steps:

[0053] The low-purity urinary follicle-stimulating hormone or menopausal gonadotropin (HMG) is purified by chromatography to obtain high-purity urinary FSH; the chromatography is cation exchange chromatography and dye affinity chromatography.

[0054] Preferably, the method comprises the steps of:

[0055] (1) purifying solution 1 containing low-purity FSH through cation exchange chromatography to obtain distillate 1; and

[0056] (2) The solution 2 containing distillate 1 was purified by dye affinity chromatography to obtain high-purity urinary follicle-stimulating hormone (pFSH).

[0057] The skeleton medium of the chromatographic column of the cation-exchange chromatography used in the preparation method of the present invention comprises cross-linked products of agarose, dextran, cellulose, styrene, acrylic acid and / or derivatives; the chromatographic column of the ...

Embodiment 1

[0099] Preparation of High Purity Urofollicle Stimulating Hormone I

[0100] Low-purity urinary follicle-stimulating hormone (purchased from Shanghai Tianwei Biopharmaceutical Co., Ltd.) was used as the starting material, wherein the biological potency of FSH was 315 IU / mg, and the biological potency of LH was ≤3 IU / mg.

[0101] Dissolve 10 g of the above-mentioned low-purity urinary follicle-stimulating hormone with 300 mL of equilibrium solution (0.03 M sodium dihydrogen phosphate, pH 5), and then put it on a 250 mL CM-Sepharose chromatography column (provided by Amersham), which has been previously used with the same equilibrium solution Well balanced. After sample loading, wash 10 times the column volume with washing solution (0.1M sodium acetate, pH5), then perform 0-100% linear gradient elution with washing solution and eluent (0.1M sodium acetate+1M NaCl, pH5), Monitor at 280nm with a UV detector, distribute and collect each distillate peak, detect its FSH immune titer...

Embodiment 2

[0107] Preparation of high-purity urinary follicle-stimulating hormone II

[0108] Dissolve 5 g of low-purity urinary follicle-stimulating hormone (the starting material is the same as in Example 1) with 150 mL of equilibrium solution (0.03 M sodium dihydrogen phosphate, pH 4.8), and then put it on a 130 mL SP-Sepharose chromatography column (provided by Amersham) , this column has been equilibrated with the same equilibrium solution in advance. After sample loading, wash 10 times the column volume with washing solution (0.1M sodium acetate, pH4.8), then perform 0-100% linear gradient washing with washing solution and eluent (0.1M sodium acetate+1M NaCl, pH5) Remove (the volume ratio concentration of the eluent, within 2 hours), monitor the 280nm place with an ultraviolet detector, distribute and collect each distillate peak, detect its FSH immune titer, combine about 0.2L of active ingredients, add pre-cooled non-toxic Precipitate with water and ethanol overnight, collect th...

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Abstract

The present invention provides a type of highly purified follicle stimulating hormone from urea, the purity of which is not less than 95w / w%, and the impurity content of which is not more than 5w / w%. And the present invention provides a method for preparing the said highly purified follicle stimulating hormone.

Description

technical field [0001] The invention relates to the fields of protein purification and biomedicine. Specifically, the present invention relates to high-purity follicle-stimulating hormone (FSH), a preparation method thereof, and a pharmaceutical composition containing it. Background technique [0002] Follicle-stimulating hormone (FSH) is a hormone produced by the pituitary gland, which consists of two subunits, the α chain and the β chain. The α subunit of FSH is exactly the same as the α subunit of luteinizing hormone (LH) and chorionic gonadotropin (CG), with 92 amino acids and a molecular weight of about 14500D, at positions 52 and 78 The asparagine on is an amino acid that undergoes N-glycosylation. [0003] The β subunit of FSH consists of 111 amino acids with a molecular weight of about 18,000 D, and the asparagine at the 7th and 24th positions are N-glycosylated amino acids. The β subunit of LH consists of 121 amino acids with a molecular weight of about 14800D; t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/59C07K1/22C07K1/18A61K38/24A61P15/08
CPCC07K14/59A61P15/08
Inventor 季晓铭高霄梁季斌郭照晔洪云海
Owner SHANGHAI TECHWELL BIOPHARMACEUTICALS CO LTD